Objective To explore the feasibility of hand-assisted laparoscopic splenectomy and azygos-portal disconnection. Methods Hand-assisted laparoscopy was performed in 12 patients with hypersplenia secondary to post-hepatitic hepatocirrhosis and a history of rupture and bleeding of esophago-gastric varicose vein.An ultrasound knife was used to dissect the ligaments of the spleen.The Endo-Cutter was used to cut off the pedicle of the spleen.Then the spleen was removed in a plastic bag.All of the varicose vessels around the fundus and the lower segment of the esophagus(6~8 cm in length) were dissected and disconnected according to the criteria of open surgery.Results The operation was successfully completed in 10 patients,while conversions to open surgery were required in 2 patients because of massive hemorrhage during the operation.The operating time was 2.5~5 h(mean,3.4 h) and the hemorrhagic volume was 100~500 ml(mean,250 ml).Postoperatively,1 patient experienced an intraperitoneal hemorrhage and received open surgery for hemostasis while the remaining patients had an uneventful recovery without complications.A total of 10 patients were followed for 0.5~2 years(mean,1.5 years).Four patients died of liver failure.Six patients presented small volumes of relapsed upper gastrointestinal bleeding around 1 year after operation.Gastroscopy showed portal hypertensive gastropathy in 3 patients,gastric ulcer in 1 patient,and ruptured varicose esophageal veins in 2 patients.All the 6 patients were cured by conservative medical treatment.Conclusions Hand-assisted laparoscopic splenectomy and azygos-portal disconnection is a feasible,effective,and safe surgical procedure.

Objective To explore the variations of regeneration hormones after hepatectomy for liver cancer,and evaluate the relationship between the liver regeneration hormones and cancer recurrence.Methods The clinical data of 129 patients with primary hepatic carcinoma in our hospital from Dec 2004 to Dec 2005 were collected.The patients were divided into three groups according to their recurrent times,which were one-month recurrence group,6 months recurrence group and one-year recurrence group.And at the same time,40 cases of liver cancer that received TAE treatment were as contrast group.Serum HGF value was detected before operation and 1,3,7,10 and 14 days after operation.c-met,which is the receptor of HGF,was also detected as c-met mRNA and protein expression in cancer tissue and near-carcinoma liver tissue by semi-quantitative RT-PCR and Western Blot.The differences between the level of expression and the time of recurrence were compared,and the results were also compared with pathological indexes.Results Serum HGF value elevated after hepatectomy,the crest time appeared at about 10 days after the operation,and decreased after 14 days.The elevated values of HGF in large HCC tumors were markedly higher than those in small HCC tumors.The change of c-met mRNA and protein levels,revealed that the earlier the recurrence in both large and small HCC,the higher the c-met levels,and the higher the rate of vascular cancer emboli.Conclusions There is marked elevation of HGF level after hepatectomy in patients with liver carcinoma,and the over expression of c-met of the tumor may be related to its early postoperative recurrence.

Objective:Cholangioduodenal reflux is still an unresolved problem in interposed jejunal choledochoduodenostomy (IJC) yet.To settle this problem,we designed IJC with distal suspending (IJCDS).Methods:IJCDS is to insert and suspend the distal end of the 25 cm-ling interposed jejunum to the duodenal cavity,which is about 2.5 cm long.The pressure of the biliary tract and the biochemical composition or the bile were measured during and after the operation and the results were compared.Barium meal and cholangiography through T tube were performed after the operation.Follow-up surveys were also done.Results:The pressure or the biliary tract decreased distinctively after the operation.Barium meals found no cholangioduodenal reflux although the patients had changed many postures.Cholangiography through T tube showed parisaltic or the interposed jejunum and the contrast medium was continuously pushed into the duodenum.Noreflux cholangitis and stone recurrence were found in the follow-up survey in 14 patients.Conclusion:IJCDS has the powerful function on preventing cholangioduodenal reflux.On the other hand,it avods the shortcomings of other reflux-preventing operative pattern.So it had the good prospect to extend its clinical usage.

Objective To investigate the effect of low molecular weight heparin (LMWH) on the expression of endothelial protein C receptor (EPCR) and protease activated receptor 1 (PAR1) in abdominal vascular endothelial cells (VECs) of septic rats. Methods VECs were cultured by tissue-sticking method, and the purity was determined with flow cytometry (FCM). VECs were randomly divided into three groups: control group, septic group (LPS 1 μg/ml) and LMWH group (LPS 1 μg/ml+LMWH 5 μg/ml). The VECs were collected at 1st, 3rd, 5th days after stimulated. The expression of EPCR and PAR1 were assessed by FCM. Results The expression of EPCR and PAR1 of septic group decreased significantly compared with control group at each time point (P＜0.05 or P＜0.01), and the expression decreased most obviously on day 5 (26.53±7.21 vs 39.26±2.62,q＝6.45,P＜0.01;53.21±15.10 vs 86.54±11.34,q＝6.94,P＜0.01). In LMWH group, the levels of EPCR and PAR1 expression were higher than setpic group at each time point (P＜0.05). Compared to control group, the expression of EPCR had a significantly decrease on day 1 (40.86±1.63 vs 45.41±2.82,q＝3.51,P＜0.05), which had no significantly different on day 3 and 5 (41.20±3.32 vs 42.83±2.66,P＞0.05;39.23±3.33 vs 39.26±2.62,P＞0.05), and the expression of PAR1 were not significantly decrease compared with control group at each time point (P＞0.05). Conclusions LMWH could improve the inhibition status and the expression of EPCR and PAR1 on VECs in septic rats.

Objective To explore the clinical effects of unilateral placement of plastic stents by ERCP for nonresectable Bismuth type IV hilar cholangiocarcinoma.Methods A prospective study was conducted in 42 patients with nonresectable Bismuth type IV hilar cholangiocarcinoma,who had unilateral insertion of plastic stents in the recent 4 years,All of the patients had successful insertion of single plastic stent by ERCP manipulation.Early results (less than 30 days) of procedure-related complications,mortality,long-term results (greater than 30 days) and survival were observed.Results All of the 42 patients had successful drainage.Mean total bilirubin value decreased from (332.3?163.4)?mol/L to (30.6?18.5)?mol/L,and complete resolution of jaundice was achieved in 61.9% (26/42)of the patients.Early comlications included papilla bleeding due to EST in 4 of 42 patients (9.5%) and acute cholangitis in 10 of 42 patients (23.8%).No procedure-related death occurred.All of the patients needed to have their stents replaced periodically,and the median duration up to the first stent replacement was 65 d,and without significant differences between the two groups in stent insertion to the left and right hepatic duct.Median duration of stent patency was 5.15 months,and median patient survival was 6.5 months,but also no significant differences were found between the two groups.Conclusions Unilateral plastic stent insertion is safe and feasible,can achieve adequate drainage for relatively long time,and can improve life quality and survival for patients with nonresectable hilar cholangiocarcinoma.

Objective To determine the allelic frequency distribution and genetic parameters of nine non-CODIS DNA index systems of the short tandemrepeat (STR ) loci (D2S1772, D6S1043, D7S3048, D8S1132, D11S2368, D12S391, D13S325, D18S1364, and GATA198B05). Methods A total of 353 blood samples were collected, extracted, amplified, and analyzed fromunrelated healthy individuals of Han na-tionality in Hunan Province, China. Results O ne hundred and fourteen alleles were observed in the pop-ulation with corresponding allelic frequencies ranged from0.001 0 to 0.323 0. For all the nine non-CODIS STR loci, the observed genotypic data showed no significant deviations fromthe Hardy-W einberg equi-librium. The Ho, He, PIC, D P, and PE of the studied non-CODIS STR loci ranged from0.108 0 to 0.195 0, 0.805 0 to 0.892 0, 0.770 0 to 0.860 0, 0.925 0 to 0.966 0 and 0.607 0 to 0.780 0, respectively. Conclusion N ine non-CODIS STR loci have high degrees of polymorphisms, which may be useful in in-dividual forensic identification and parentage testing in forensic practice.

Objective To identify the surface marker of bone marrow-derived liver stem cells and to isolate the stem cells, to investigate the differentiation of the stem cells. Methods The quantitative variations of the cells with stem cell surface markers, including ? 2-microglobulin negative (? 2m -), Thy-1 +,CD34 +,Flt-3 +,IL-3R +,and c-kit + markers, were detected by using flow cytometry in the bone marrow of several rat models with liver injury. Each stem cell population was then isolated using a magnetic bead cell-sorting procedure. The isolated cells were cultured in a system containing cholestatic serum and hepatocyte growth factor (HGF). The morphology of the cells was observed, and the expressions of albumin, AFP, and CK8/18 were detected with immunohistochemistry technique. Results ? 2m - cells elevated significantly in each of the rat models.After being co-cultured with cholestatic serum and HGF, ? 2m - cells showed multilateral transformation and resembled hepatcytes morphologically. The differentiated cells expressed albumin, AFP, and CK8/18, all known to be the characteristic markers of hepatocyte. The other cell population showed little quantitative changes, and did not express the same proteins. Conclusions The quantitative variation of ? 2m - cells corresponds to the severity of liver injury. ? 2m - cells have the ability to trans-differentiated into hepatocytes in vitro. They might be the marker of liver stem cells.

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro . METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.

Objective To investigate the presences of regulatory T cells (Treg) and natural killer T cell (NKT)-like T cells in the peripheral blood of healthy subjects and patients with newly diagnosed cancer. Methods Patients were enrolled into four groups: healthy subjects (Group A), patients without cancer (Group B), patients with newly diagnosed stage Ⅰ~Ⅱ cancer (Group C), and patients with newly diagnosed stage Ⅲ~Ⅳ cancer (Group D). Flow cytometry was performed to detect the percentage of CD4+ CD25high CD127low cells in CD4+ T lymphocytes (Treg) and the percentage of CD3+ CD56+ cells in CD3+ T lymphocytes (NKT-like). Results The percentage of Treg cells in CD4+T lymphocytes was(6.72 ± 3.16)%, (6.49 ± 2.83)%,(6.80 ± 3.07)%and (7.63 3.47)% in Group A, B, C and D, respectively, with no significant differences among these groups (P>0.05). However, the percentage of CD3+CD56+T cells in CD3+T lymphocytes was (3.13 ± 1.66)%, (2.56 ± 1.27)%,(7.22 ± 2.70)% and (7.72 ± 3.10)% in Group A, B, C and D, respectively, with marked increases in Group C and D compared to those in Group A and B (P < 0.05). Conclusion Patients with newly diagnosed cancer demonstrated increasing trends in the percentage of peripheral NKT-like cells.

This article introduces the basic theories about atomic force microscope (AFM) and electron microscope (EM), respectively. New applications of each microscopic technology in regenerative medicine, covering both material science and life science, are discussed. The advantages or disadvantages of the kinds of microscopes in working conditions, sample preparation, resolution and the like, are discussed and compared systematically to make clear each scope of applications. This could be a useful guide for selecting the appropriate microscopic analysis in research work about regenerative medicine.
Humans ; Microscopy, Atomic Force ; methods ; trends ; Microscopy, Electron ; methods ; trends ; Regenerative Medicine ; methods ; trends